Herpes simplex virus type 2 (HSV-2) infects a half-billion individuals worldwide, which has important public health implications since HSV-2 genital ulcer disease increases the risk of acquiring and transmitting HIV by 3-fold. Our work focuses on immune evasion strategies of HSV-2 glycoproteins C (gC2) and E (gE2). HSV-2 gC2 binds complement component C3b to inhibit complement activation. HSV-2 gE2 blocks Fc- mediated activities by binding the Fc domain of an IgG antibody molecule that is bound by its F(ab')2 domain to HSV antigen, which inhibits complement activation and antibody-dependent cellular cytotoxicity (ADCC). In Aim 1 we will evaluate the hypothesis that gC2 and gE2 immune evasion greatly reduce the effectiveness of an HSV-2 glycoprotein D (gD2) vaccine in humans by inhibiting antibody and complement, and that gC2 and gE2 immune evasion contribute to recurrent genital disease after naturally acquired infection. We will test our hypothesis by evaluating sera from subjects immunized with the GSK gD2 subunit vaccine to determine whether gC2 and gE2 immune evasion inhibit antibody and complement neutralization and ADCC, and by assessing sera obtained from subjects with multiple recurrences of genital HSV-2 or with no recurrences to evaluate the contribution of gC2 and gE2 immune evasion to recurrent genital ulcer disease. In Aim 2 we will assess the hypothesis that HSV-2 gC2 and gE2 mutant strains defective in immune evasion will be log10 orders of magnitude less virulent than wild-type or recue virus in mouse and guinea pig vaginal infection models. We will test our hypothesis by constructing single gC2 and gE2 mutant strains and a double gC2/gE2 mutant strain that are defective in immune evasion but intact for replication, protein expression and other functions ascribed to them. We will determine the magnitude of the impact of immune evasion in mice and guinea pigs and define mechanisms using complement and NK deficient animals. In Aim 3 we will evaluate the hypothesis that a gC2/gD2/gE2 vaccine totally protects against genital ulcer disease and markedly reduces or totally prevents latency. We will define the optimum dose of gC2 and gE2 antigen that induce high titers of antibodies that block immune evasion and of gD2 antigen that induces high titers of neutralizing antibodies. We will then evaluate whether the trivalent vaccine provides better protection against genital disease and latent infection than any single antigen or double antigen combination. To improve the translatability of animal models to human studies, we will modify endpoint determinations in animal models to match human trials, including confirming disease scores by seroconversion, PCR and culture. We will determine if local virus replication in vaginal tissues is sufficient to induce seroconversion in immunized animals that have no genital disease or latent infection. Detecting seroconversion under these conditions would be informative since it is may not serve as a useful marker for latency in human trials. These studies take a novel approach to vaccine development and may lead to a new generation of vaccines.